Video 8: Neuronal activity traces of representative neurons.

A visualization of the neuronal traces shown in Figure 7b of the related publication; presenting rapid volumetric calcium imaging of highly motile Hydra.<div><br><div>An advantage of the LSTM imaging technique (see below for details) is the capacity for rapid volumetric live imaging of samples undergoing substantial non-isomorphic rearrangements in their body shape and cellular density, resulting in continuously changing local optical properties.<br><div><br></div><div>This video relates to the development of the imaging technique LSTM: Light Sheet Theta Microscopy for rapid high-resolution imaging of large biological samples, an approach designed to overcome the limitations of 'standard' Light Sheet microscopy LSM in terms of lateral dimensions and imaging quality, while preserving the benefits of LSM in terms of imaging resolution, depth and speed.</div><div><br></div><div>LSTM uses two symmetrically-arranged oblique light-sheets, generated using independent illumination objectives, for rapid high-resolution imaging of large samples.</div><div><br></div><div>View the full collection of related videos at: </div><p><a href="https://doi.org/10.6084/m9.figshare.c.4072160">https://doi.org/10.6084/m9.figshare.c.4072160</a></p></div></div

Video 5: Visualization of an image stack of vasculature stained rat brain tissue.

This video visualizes an image stack acquired from a large rat brain slice (stained for vasculature with tomato lectin) using LSTM in 2-AS (2-axes scan) mode. The bounding box is 1mm x 1mm x 5mm. Uniform high-resolution imaging of the entire tissue under LSTM is presented.<div><div><br></div><div>This video relates to the development of the imaging technique LSTM: Light Sheet Theta Microscopy for rapid high-resolution imaging of large biological samples, an approach designed to overcome the limitations of 'standard' Light Sheet microscopy LSM in terms of lateral dimensions and imaging quality, while preserving the benefits of LSM in terms of imaging resolution, depth and speed.</div><div><br></div><div>LSTM uses two symmetrically-arranged oblique light-sheets, generated using independent illumination objectives, for rapid high-resolution imaging of large samples.</div><div><br></div><div>View the full collection of related videos at: </div><p><a href="https://doi.org/10.6084/m9.figshare.c.4072160">https://doi.org/10.6084/m9.figshare.c.4072160</a></p></div

Video 1: 3D model of LSTM implementation.

<div>The 3D modelling and rendering was performed using Autodesk Inventor 2017, and the animation was performed using Autodesk Fusion 360 2017 and MATLAB. The components labelled are LS (laser source), collimator, ND (neural density) filter mount, iris, ETL (electrically tunable lens), slit, CL (cylindrical lens), Galvo scanner, SL (scan lens), iris and TL (tube lens).</div><div><br></div><div>The LSTM illumination and detection arms are implemented as rigid assemblies (using the caging system from Thorlabs), connected to a vertically mounted breadboard via x-y manual translation stages for finer adjustments.</div><div><div><br></div><div>This video relates to the development of the imaging technique LSTM: Light Sheet Theta Microscopy for rapid high-resolution imaging of large biological samples, an approach designed to overcome the limitations of 'standard' Light Sheet microscopy LSM in terms of lateral dimensions and imaging quality, while preserving the benefits of LSM in terms of imaging resolution, depth and speed.<br></div><div><br></div><div>LSTM uses two symmetrically-arranged oblique light-sheets, generated using independent illumination objectives, for rapid high-resolution imaging of large samples.</div><div><br></div><div>View the full collection of related videos at: </div><p><a href="https://doi.org/10.6084/m9.figshare.c.4072160">https://doi.org/10.6084/m9.figshare.c.4072160</a></p></div

Video 7: Rapid volumetric calcium imaging of highly motile Hydra.

GCaMP6s expressing Hydra was imaged using LSTM with 10x/0.6NA objective. Maximum intensity projections are shown for the two halves of the volume. First occurrences of longitudinal and radial contractions are annotated. The scale Bar is 100 microns.<div><br></div><div>An advantage of the LSTM imaging technique (see below for details) is the capacity for rapid volumetric live imaging of samples undergoing substantial non-isomorphic rearrangements in their body shape and cellular density, resulting in continuously changing local optical properties.<div><div><br></div><div>This video relates to the development of the imaging technique LSTM: Light Sheet Theta Microscopy for rapid high-resolution imaging of large biological samples, an approach designed to overcome the limitations of 'standard' Light Sheet microscopy LSM in terms of lateral dimensions and imaging quality, while preserving the benefits of LSM in terms of imaging resolution, depth and speed.</div><div><br></div><div>LSTM uses two symmetrically-arranged oblique light-sheets, generated using independent illumination objectives, for rapid high-resolution imaging of large samples.</div><div><br></div><div>View the full collection of related videos at: </div><p><a href="https://doi.org/10.6084/m9.figshare.c.4072160">https://doi.org/10.6084/m9.figshare.c.4072160</a></p></div></div

Video 2: Comparison of image volumes acquired with LSTM in 1-AS and 2-AS modes.

<div><p>The 3D rendering visualizes the image stacks acquired from the same sample (human brain section stained with DAPI) with LSTM in 1-Axis Scan (1-AS) and simultaneous 2-Axes Scan (2-AS) mode.</p><p><br></p> <p>This video relates to the development of the imaging technique LSTM: Light Sheet Theta Microscopy for rapid high-resolution imaging of large biological samples, an approach designed to overcome the limitations of 'standard' Light Sheet microscopy LSM in terms of lateral dimensions and imaging quality, while preserving the benefits of LSM in terms of imaging resolution, depth and speed.</p></div><div><br></div><div>LSTM uses two symmetrically-arranged oblique light-sheets, generated using independent illumination objectives, for rapid high-resolution imaging of large samples.</div><div><br></div><div>View the full collection of related videos at: </div><p><a href="https://doi.org/10.6084/m9.figshare.c.4072160">https://doi.org/10.6084/m9.figshare.c.4072160</a></p

Video 4: High-resolution LSTM imaging of a large tissue of Thy1-eYFP mouse brain.

The bounding box is 9.6mm x 13.5 mm x 5.34 mm. The raw data was down-sampled 4x4 fold to make the volume rendering feasible.<div><br></div><div>Imaging presents a thick coronal slice (~9.6mm x 13.5 mm x 5.34 mm; the sample expanded ~1.5-2 fold due to the immersion in glycerol solution) of a CLARITY-cleared <i>Thy1-eYFP</i> transgenic mouse brain, with 10x/0.6NA/8mmWD.</div><div><br></div><div>This video relates to the development of the imaging technique LSTM: Light Sheet Theta Microscopy for rapid high-resolution imaging of large biological samples, an approach designed to overcome the limitations of 'standard' Light Sheet microscopy LSM in terms of lateral dimensions and imaging quality, while preserving the benefits of LSM in terms of imaging resolution, depth and speed.</div><div><br></div><div>LSTM uses two symmetrically-arranged oblique light-sheets, generated using independent illumination objectives, for rapid high-resolution imaging of large samples.</div><div><br></div><div>View the full collection of related videos at: </div><p><a href="https://doi.org/10.6084/m9.figshare.c.4072160">https://doi.org/10.6084/m9.figshare.c.4072160</a></p